• The Journal of Korean Society of Virology
• /
• v.29 no.4
• /
• pp.247-259
• /
• 1999

Small round structured viruses (SRSV) are the major ethological agents which can cause outbreaks of non-bacterial gastroenteritis or food poisoning both in children and adults. The classification of family Caliciviridae to which SRSV belong, is based on the genome encoding three open reading frames. The rotavirus is another major pathogen which causes diarrhea in young children. We examined stool specimens obtained from diarrheal patients in Wonju from which bacterial pathogens were not found. To detect causative viruses from stool specimens of patients, reverse transcription (RT)-polymerase chain reaction (PCR) or nested PCR using rotavirus or SRSV specific primers was performed. In this study, RT-nested PCR procedure which can amplify a 330 bp fragment derived from RNA dependent RNA polymerase (RDRP) region within ORF1 was applied for the detection of SRSV. For the detection of rotaviruses, a 877 bp fragment from the VP4 region of rotavirus genome was amplified. As a result, rotavirus was not detected while SRSV sequences were detected from one out of five specimens. The nucleotide and amino acid sequences of the Wonju isolate were compared with other 6 Korean isolates which have been isolated and sequenced in our laboratory. Sequence analysis revealed that the Wonju isolate was rather distinct from other Korean isolates: the Wonju isolate was closer to genogroup I of SRSV while other 6 Korean isolates belonged to genogroup II.

• Microbiology and Biotechnology Letters
• /
• v.43 no.3
• /
• pp.296-299
• /
• 2015

Cowpea mild mottle virus (CPMMV) has a wide range of hosts, such as the pea family and tomato. CPMMV is a non-reported virus in Korea, and is domestically designated as a controlled virus associated with plant quarantine. In this study, a rapid diagnostic method for the detection of CPMMV at quarantine sites was developed. For the development of a user-based system, the PCR compositions and conditions use existing methods of quarantine for the viruses. Two sets of RT-PCR and nested PCR were developed in this study that could be amplified from 579 → 298 dp and 638 → 252 bp, respectively. Furthermore, a sequence inserted positive control plasmid was developed, which is able to identify false-positives resulting from laboratory contamination. The findings of this study are important for the diagnosis of CPMMV in imported crops held in plant quarantine.

• Journal of Life Science
• /
• v.21 no.6
• /
• pp.819-828
• /
• 2011

To inspect norovirus contamination of groundwater in south eastern areas of Korea, a systematic survey of groundwater in Busan, Ulsan, and Gyeongsangnam-do was performed for two years from 2009 to 2010. For this purpose, we first optimized the nested reverse transcription-PCR condition by designing two sets of primers for the detection of norovirus genogroups, GI and GII. Of 145 samples, 21 (25.9%) and 15 (23.4%) were norovirus positive in the dry season (April to June) and wet season (July to August), respectively. The detection frequencies of norovirus in Busan, Ulsan, and Gyeongsangnam-do were 15%, 7%, and 32%, respectively, reflecting a geographical difference in their distribution. The GI and GII isolates were 5 and 31, respectively, indicating the prevalence of GII in the tested areas. According to phylogenetic analysis of their nucleotide sequences, all of the GI isolates were identified to genotype GI.5 whilst the GII isolates were divided into two genotypes, GII.3 and GII.4. Neither physical-chemical parameters such as pH, temperature, oxidation-reduction potential, and dissolved oxygen, nor microbial indicators of water quality such as total bacteria, total coliforms, and Escherichia coli were statistically correlated with contamination of norovirus in the groundwater. Interestingly, however, the presence of norovirus was closely correlated with low turbidity (<0.50 NTU). The present study suggests that periodical monitoring of norovirus in groundwater is necessary to prevent epidemic waterborne diseases and to secure better sanitary conditions for public health.

• The Plant Pathology Journal
• /
• v.31 no.4
• /
• pp.438-440
• /
• 2015

We developed a loop-mediated isothermal amplification (LAMP) method to rapidly diagnose Wheat streak mosaic virus (WSMV) during quarantine inspections of imported wheat, corn, oats, and millet. The LAMP method was developed as a plant quarantine inspection method for the first time, and its simplicity, quickness, specificity and sensitivity were verified compared to current reverse transcription-polymerase chain reaction (RT-PCR) and nested PCR quarantine methods. We were able to quickly screen for WSMV at quarantine sites with many test samples; thus, this method is expected to contribute to plant quarantine inspections.

• Korean Journal of Clinical Laboratory Science
• /
• v.52 no.2
• /
• pp.143-149
• /
• 2020

The study examined the seasonal pattern of larvae, nymph, and adult life stages for Haemaphysalis longicornis and the rate of infection with severe fever from the thrombocytopenia syndrome virus in ticks collected from 12 sections (Jichukdong), 14 sections (Uldaeri), and 18 sections (Howondong) in Bukhansan dullegil for April-October 2019. Haemaphysalis longicornis ticks have been considered the main vector for severe fever with thrombocytopenia syndrome (SFTS). Haemaphysalis flava and Ixodes nipponensis were collected using the dragging and flagging method. The ratios of Haemaphysalis longicornis of the collected ticks were 91% (Jichukdong), 94% (Uldaeri), and 98% (Howondong). Monthly distributional studies of Haemaphysalis longicornis based on the developmental stage showed that the adults peaked in September while nymphs were collected more frequently from April through June. The larvae peaked in September and October. SFTS virus detection was performed using 2 × OneStep RT-PCR and nested PCR. On the other hand, no SFTS virus-specific gene was detected in 1,158 ticks of Haemaphysalis longicornis. This result provides estimates of the population densities for the life stages of Haemaphysalis longicornis and the associated disease risk in Bukhansan dullegil, where many people have visited since opening in 2010.

• The Journal of Korean Society of Virology
• /
• v.29 no.1
• /
• pp.45-53
• /
• 1999

We examined the hepatitis G virus infections among 227 Koreans who were healthy or were suspected of hepatitis and determined the phylogenetic relationship based on a part of the NS-5 region of 5 positive samples. Viral RNA was extracted from sera and cDNA was synthesized and subsequently amplified by RT-PCR (reverse transcription-polymerase chain reaction) or RT-nested PCR using random hexamer and NS-5 specific primers (470-20-1-77F, 470-20-1-211R, HGVNESTFO, HGVNESTRE). Five positives were found to belong to samples of patients showing symptoms of viral hepatitis. Primers used for PCR or nested PCR were derived from the NS-5 region. On the other hand, no amplification was detected using primers derived from the 5'-NCR (G-146F, G-401R). We performed TA cloning and sequencing of 5 amplified fragments, and their sequences were compared with those of foreign isolates of HGV. The phylogenetic analysis using MegAlign programme of DNAstar has shown that the Korean isolates are clustered on the phylogenetic tree. In summary, we confirmed the hepatitis G virus infection in 5 cases out of 12 patients showing the symptoms of viral hepatitis. The phylogenetic analysis of sequences of 5 amplified fragments showed that their relations to each other were closer than those to the foreign HGV isolates reported.

• Biomedical Science Letters
• /
• v.28 no.2
• /
• pp.83-91
• /
• 2022

This study was performed to investigate the contamination status of microorganisms that cause food poisoning in shellfish distributed in the metropolitan area of South Korea. The analyses were conducted according to the sample type, season, and region. In particular, the genotype of food poisoning viruses that explosively cause collective food poisoning was analyzed. Total 483 each of four types of shellfish (Crassostrea gigas, Mytilus coruscus, Pectea albicans albicans, and Scapharca subcrenata) were collected from four distribution sites located in the metropolitan area. We investigated foodborne pathogen by multiplex PCR and RT/Nested PCR from shellfish. As a result, Vibrio parahaemolyticus and Bacillus cereus were detected in 13 and 21 samples and Norovirus (NoV) GI and GII were detected in 7 and 12 samples, respectively. V. parahaemolyticus and NoV GI and GII showed differences in types of shellfish and seasons, and no correlation was confirmed with regional differences. Also, as a result of genotyping for the detected NoV GI and GII, they were confirmed as NoV GI.7, GI.5 and GII.3, which are causes of collective food poisoning. Therefore, cross-infection with shellfish can cause food poisoning. In particular, attention must be paid to the handling and cooking of shellfish in organizations that implement group feeding, and it is necessary to establish a management system for microorganisms that cause food poisoning in the process of shellfish farming and distribution.

• Tuberculosis and Respiratory Diseases
• /
• v.53 no.3
• /
• pp.265-274
• /
• 2002

Background : Tumor associated antigens, which are produced specifically by tumor cells, are promising targets for the early diagnosis and immunotherapy. Among the tumor associated antigens, MAGE (a melanoma antigen), BAGE, GAGE, PRAME and NY-ESO were named as cancer/testis specific antigens they are detected exclusively in the testis or cancer cells If MAGE is easily detectable in the sputum, it would become a convenient method for diagnosing lung cancer. This study was undertaken to investigate MAGE expression in the induced sputum obtained from lung cancer patients. Materials and Methods : In 14 control patients and 30 lung cancer patients, the induced sputum was collected after inhaling 3% saline(5 cc) delivered by nebulizer for approximately 5 minutes after a mouth rinse and bronchodilator inhalation. The induced sputum was placed in a conservative-mixed solution (guanidinium isothiocyanate, Triton X-100). The total cellular mRNA was extracted from the cells and RT PCR and nested PCR were run in 30 and 35 cycles respectively, with two different types of primers specially designed to detect six subtypes of MAGE DNA simultaneously. Results : MAGE expression was not detected in the 14 controls, but in the 30 cancer patients, MAGE was found in 24 patients (80%, p=0.001). In the cancer patients, there were no differences in the expression level according to the tissue types (squamous cell cancer 13/17, adenocarcinoma 7/9, and small cell cancer 4/4, p-0.56). Among the 24 MAGE-positive patients, the tumor was not visible on a bronchoscopy in 11 patients (45.8%). Conclusion : A study of MAGE in induced sputum appears to be a useful and complementary method in the diagnosis of lung cancer. A further prospective study with more patients is recommended.

• Journal of fish pathology
• /
• v.28 no.1
• /
• pp.27-35
• /
• 2015

VHSV is a major viral agent that affects freshwater and marine fish, causing serious economic losses in aquaculture in the world. Due to their filter-feeding activity, bivalve mollusks may act as viral transmitters after accumulation of the fish viruses released into seawater from infected fish. Amplification by RT-PCR was carried out to investigate the presence of VHSV in pacific oysters (Crassotrea gigas) and blue mussels (Mytilus edulis), inhabiting regions around aquatic farms in Korea. Primers designed from conserved regions of VHSVs allowed us to detect four different types of VHSV in a single PCR. Twenty two of the eighty four samples showed positive results of VHSV in a 2-step RT-PCR. Using six positive samples from three different regions in Korea, we cloned and sequenced the glycoprotein (G) gene (467-bp long) of VHSVs. Genetic analysis of the VHSVs detected in shellfish in various geographical areas of Korea showed highly restricted results to VHSV type Iva. This was in agreement with the reports showing only a single genotype of VHSV (Iva genotype) in outbreaks in cultured or wild fish in Korea. Consequently, we investigated VHSVs carried by bivalve mollusks inhabiting the vicinity of aquatic farms, and revealed correlationship between the type of viral accumulated in shellfish by filter-feeding, and those detected in disease outbreaks in fish.

• Korean Journal of Microbiology
• /
• v.49 no.2
• /
• pp.112-117
• /
• 2013

Wheat streak mosaic virus (WSMV), a member of the genus Tritimovirus in Potyviridae, severely impacts wheat and corn seed worldwide, but has yet to be detected in Korea, and hence, every effort should be made to prevent its introduction. To prevent WSMV from entering the country, it is necessary to prepare a specific, sensitive, simple, and fast detection method for routine application to plant quarantine procedures. For this reason, a two-step diagnosis system consisting of RT-PCR and nested PCR is being used for WSMV detection. In addition, a novel positive control was developed for use with the system. WSMV has been detected in seed sweet corn from Japan and seed wheat from USA by a two-step diagnosis system, the details of which are described in this study. After sequence analysis, similarities of 80.6 and 100.0% with other isolates were determined by BLAST. They showed the same topology, which was classified as 4 genotypes by various phylogenetic trees, using a poly protein encoding sequence amplification. In this analysis, WSMV-JSweet-corn2868 (JX845574) is classified as clade B, while WSMV-Uwheat1944-1 (KC754959) and WSMV-Uwheat1944-2 (KC754960) belong to clade D.